Cd45+ Cell Analysis
Cd45+ Cell Analysis - Related Videos
16:55
Isolation & Characterization of Hoechstlow CD45negative Mouse Lung Mesenchymal Stem Cells
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Cited by 13 •
2011
In this article we demonstrate the isolation of murine resident lung mesenchymal stem cells (lung MSC), their expansion, characterization and analysis of immunomodulatory properties.
12:14
Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain
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Cited by 80 •
2016
Inflammation plays a central role in the pathogenesis of ischemic stroke. Increasing evidence suggests that it acts as a double-edged sword which exacerbates early brain injury, but also contributes to later repair. This protocol describes the isolation of immune cells from the ischemic brain and their subsequent flow cytometric phenotyping.
04:38
Analysis of Immune Cells in Mouse Nervous Tissues Using Flow Cytometry
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2025
Source: Hidmark, A. S., et al. Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry. J. Vis. Exp. (2017). This video demonstrates the use of flow cytometry to analyze immune cells in mouse sciatic nerves and dorsal root ganglia (DRGs). The process involves labeling immune cells with fluorophore- and biotin-tagged antibodies, stabilizing and permeabilizing the cells, and using fluorescent markers to detect cell-surface and nuclear...
11:08
Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry
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Cited by 4 •
2017
This article describes spectral cytometry, a new approach in flow cytometry that uses the shapes of emission spectra to distinguish fluorochromes. An algorithm replaces compensations and can treat auto-fluorescence as an independent parameter. This new approach allows for the proper analysis of cells isolated from solid organs.
05:58
Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions
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Cited by 21 •
2016
The skin is home to a complex immune cell network. We describe an efficient methodology for the digestion of mouse skin, from different parts of the animal's body, in order to obtain a single-cell suspension and analyze the different leukocyte populations resident in the skin by flow cytometry.
FAQs
Fluorescently labeled anti-CD45 antibodies mark leukocyte-associated events so researchers can identify them during flow-cytometric analysis. Additional antibodies against lymphoid, myeloid, or other immune markers then subdivide the CD45-positive population. This layered approach converts a broad immune-cell signal into more informative population-level data for examining which leukocyte groups are present in a cancer sample.
CD45 identifies a broad leukocyte population rather than a single immune-cell type. Researchers therefore combine the CD45 signal with markers associated with lymphoid, myeloid, or other immune populations. This classification helps distinguish different cellular contributors within a tumor sample and supports more specific interpretation of immune-cell composition than CD45 detection alone.
Flow cytometry uses fluorescent labeling to detect and further classify CD45-positive events, making it suited to characterizing immune-cell populations in a sample. Immunohistochemistry instead contributes spatial information by showing where CD45-expressing leukocytes occur within tissue. Using either approach depends on whether the study emphasizes population characterization or the distribution of immune cells in the tumor environment.
CD45-based analysis separates leukocyte-associated events from most nonhematopoietic cells, which can help researchers evaluate malignant cells alongside surrounding hematopoietic populations. Adding immune-lineage markers provides further classification of the CD45-positive compartment. This distinction is valuable when interpreting the cellular makeup of tumors and determining how much of a sample reflects immune infiltration rather than the malignant population.
A typical workflow labels the sample with a fluorescent anti-CD45 antibody, detects CD45-positive events by flow cytometry, and then applies additional immune-cell markers to classify the detected populations. The resulting measurements describe the leukocyte compartment and its lymphoid or myeloid components, providing a structured basis for comparing immune composition across cancer samples or study conditions.
Immunohistochemistry is useful when the location of leukocytes within tissue matters in addition to their presence. CD45 staining can reveal the spatial distribution of immune cells relative to the tumor and surrounding tissue. That information complements population-based flow-cytometry results by showing whether immune-cell infiltration is localized, dispersed, or associated with particular tissue regions.
Researchers can use these measurements to evaluate immune-cell infiltration and characterize the tumor microenvironment. Comparing CD45-positive populations across samples can support studies of disease progression and tumor immunity. When applied across treatment conditions, the analysis can also contribute to evaluating responses to immunotherapy by showing changes in the immune-cell compartment.
The method provides a way to quantify or localize the leukocyte component surrounding malignant cells. Flow cytometry describes the composition of CD45-positive immune populations, while immunohistochemistry adds tissue-level context. Together, these observations help researchers relate immune-cell infiltration to tumor biology, disease progression, and treatment-response studies without treating the tumor as a purely malignant-cell population.